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Virologica Sinica, 19 (3) : 214-216, 2004
Research Article
The Study of Amplifing HCV ssRNA by RT-PCR Combined With Transcription in vitro
Lab of Molecular Virus & Cancer, Institute of Virology, Medical College, Wuhan University, Wuhan 430071, China
The purpose of this study is to establish a new method which HCV RNA can be used to detect sensitively and specifically through efficient single strand RNAs(ssRNAs) amplification. The sera from the chronic Hepatitis C patients were used as specimens. Single step reverse transcription-PCR (SRT-PCR) was performed prospectively, with a designed set of primers which defined the sequence unique to the 5’ noncoding region of the HCV genome, a great deal of specific ssRNAs were synthesized in the presence of T7RNA polymerase and other appropriate conditions within five hours. Comparing with the nested-PCR, the quantity of the products in our method was nearly 20-fold higher and its specificity was verified by the electrophoresis on agarose gel and the dot hybridization with the probe.
  Published online: 20 Jun 2004
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